Structural identification of recombinant human CD83 mutant variant as a potent therapeutic protein

Protein Expr Purif. 2010 Oct;73(2):140-6. doi: 10.1016/j.pep.2010.05.016. Epub 2010 Jun 8.


The formation of aberrant disulfide bonds is a structural consideration for the manufacturing of the extracellular domain of human CD83 (hCD83ext), a potential therapeutic protein. In certain instances, hCD83ext protein products, even when stored frozen, tended to dimerize or even multimerize through the formation of aberrant intermolecular disulfide bonds. Herein, we discovered an analytical inconsistency and applied a modified sample preparation protocol for proper structural analysis of hCD83ext products which are heterologously expressed in Escherichia coli and subsequently purified. In addition, a mutant derivative with the Cys100Ser mutation was identified as an improved version which did not form dimers or multimers. The identification of this mutant variant as a more potent therapeutic protein than other hCD83ext species demonstrated that the structural variation associated with disulfide bond formation can be a critical issue for rigorous control of the quality and bioactivity of therapeutic proteins. The application of this mutant variant for protein therapeutics is currently under exploration.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Antigens, CD / chemistry
  • Antigens, CD / genetics*
  • Antigens, CD / therapeutic use
  • Dimerization
  • Disulfides / chemistry
  • Disulfides / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Fermentation
  • Genetic Variation
  • Glutathione Transferase / metabolism
  • Humans
  • Immunoglobulins / chemistry
  • Immunoglobulins / genetics*
  • Immunoglobulins / therapeutic use
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / genetics*
  • Membrane Glycoproteins / therapeutic use
  • Mutagenesis, Site-Directed
  • Mutation*
  • Protein Processing, Post-Translational
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / therapeutic use
  • Serine / metabolism
  • von Willebrand Factor / genetics
  • von Willebrand Factor / metabolism


  • Antigens, CD
  • CD83 antigen
  • Disulfides
  • Immunoglobulins
  • Membrane Glycoproteins
  • Recombinant Proteins
  • von Willebrand Factor
  • Serine
  • Glutathione Transferase